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Localization of the pre-squalene segment of the isoprenoid biosynthetic pathway in mammalian peroxisomes

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Abstract

Previous studies have indicated that the early steps in the isoprenoid/cholesterol biosynthetic pathway occur in peroxisomes. However, the role of peroxisomes in cholesterol biosynthesis has recently been questioned in several reports concluding that three of the peroxisomal cholesterol biosynthetic enzymes, namely mevalonate kinase, phosphomevalonate kinase, and mevalonate diphosphate decarboxylase, do not localize to peroxisomes in human cells even though they contain consensus peroxisomal targeting signals. We re-investigated the subcellular localization of the cholesterol biosynthetic enzymes of the pre-squalene segment in human cells by using new stable isotopic techniques and data computations with isotopomer spectral analysis, in combination with immunofluorescence and cell permeabilization techniques. Our present findings clearly show and confirm previous studies that the pre-squalene segment of the cholesterol biosynthetic pathway is localized to peroxisomes. In addition, our data are consistent with the hypothesis that acetyl-CoA derived from peroxisomal β-oxidation of very long-chain fatty acids and medium-chain dicarboxylic acids is preferentially channeled to cholesterol synthesis inside the peroxisomes without mixing with the cytosolic acetyl-CoA pool.

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Abbreviations

AOX:

Acyl-CoA oxidase

FPP:

Farnesyl diphosphate

GFP:

Green fluorescent protein

HMGCR:

HMG-CoA reductase

IDI1:

Isopentenyl diphosphate isomerase 1

ISA:

Isotopomer spectral analysis

MID:

Mass isotopomer distributions

MPD:

Mevalonate diphosphate decarboxylase

MVK:

Mevalonate kinase

PEX:

Peroxin

PMP:

Peroxisomal membrane protein

PMVK:

Phosphomevalonate kinase

PP:

Diphosphate

PTS:

Peroxisomal targeting signal

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Acknowledgments

This work was supported by National Institutes of Health (NIH) grants DK58238 and DK58040 to S.K.K. We thank Drs. Eveline Baumgart-Vogt and Herbert Stangl for helpful discussions and comments on the manuscript.

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Authors and Affiliations

  1. Department of Biology, San Diego State University, San Diego, CA, USA

    Werner J. Kovacs, Khanichi N. Tape, Janis E. Shackelford & Skaidrite K. Krisans

  2. Department of Anatomy and Cell Biology, Division of Medical Cell Biology, Justus Liebig University of Giessen, Aulweg 123, 35385, Giessen, Germany

    Werner J. Kovacs

  3. Department of Nutrition, Case Western Reserve University, Cleveland, OH, USA

    Xueying Duan, Takhar Kasumov & Henri Brunengraber

  4. Department of Biomedical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA

    Joanne K. Kelleher

  5. Institute of Cell Biology, Schafmattstrasse 18, ETH-Hönggerberg, HPM, F 39.2, 8093, Zürich, Switzerland

    Werner J. Kovacs

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  1. Werner J. Kovacs
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  2. Khanichi N. Tape
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  3. Janis E. Shackelford
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Correspondence to Werner J. Kovacs.

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Kovacs, W.J., Tape, K.N., Shackelford, J.E. et al. Localization of the pre-squalene segment of the isoprenoid biosynthetic pathway in mammalian peroxisomes. Histochem Cell Biol 127, 273–290 (2007). https://doi.org/10.1007/s00418-006-0254-6

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